Rathnamma, D.Shrikrishna, IsloorVeeregowda, B.M.Chandranaik, B.M.Manjunatha Reddy, G.B.Sreenivasa Babu, T.--, U-2017-11-032017-11-032015-08-21----http://krishikosh.egranth.ac.in/handle/1/5810034710P.G. ThesisThe present study was undertaken with the objectives of isolation of Orf virus (ORFV) in cell cultures and chick embryos and detection of ORFV by PCR and standardization of LAMP assay. A total of 70 samples collected from suspected cases of Orf disease in sheep and goats, in eight districts of Karnataka from 27 outbreaks. Out of which three were organized and 24 were unorganized sheep and goats farms. The highest prevalence of disease was observed in goats compared to sheep and more deaths were seen in kids. Maximum numbers of outbreaks were reported in unorganized farms. The PCR was standardized using B2L gene for detection of ORFV. The screening of 70 samples for B2L gene amplification revealed 60 per cent positivity of ORFV. The phylogenetic analysis based on nucleotide and deduced amino acid sequences of full length and partial B2L gene revealed that, the ORFV isolates had highest genetic homology among them and with the other Indian ORFV isolates. The maximum genetic heterogeneity was observed with ORFV isolates from other countries. Further, full length B2L gene sequence based analysis provided significant information about the genetic relationship among the ORFV isolates. The ORFV was successfully isolated in the ECE by CAM route of inoculation but not in SLT and Vero cell cultures. The LAMP assay was standardized using HNB dye for the easy detection of ORFV in PCR positive scab samples. The successful amplification was indicated by a colour change from deep blue to light blue. All the PCR positive samples were also found positive with LAMP assay. The LAMP assay is rapid, easy, less expensive and most importantly, it can diagnose the disease at the point of care without thermal cycler and gel electrophoresis and is especially applicable in a resource-limited situation.ennull--Thesis